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. 2018 Jul 13;8:10602. doi: 10.1038/s41598-018-28896-8

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Detection and measurement of trapped air in ampoules using Tesla coil leak detector and air absorbing agents. (a) Tesla-negative ampoule containing trapped air. (b) Tesla-positive ampoule containing no or minimal air. (c) Percentage of Tesla-positive and Tesla-negative ampoules out of 198 and the relative proportion (%) of ampoules containing variable amounts of trapped air. Comparison of comet tail lengths of FD spermatozoa stored in (d) Tesla-positive and Tesla-negative ampoules, and (e) Tesla-treated and Tesla-untreated ampoules. (f) Result of anti-gamma-H2AX antibody staining. Brightness plots of male pronuclei obtained from fertilization of oocytes with 1-day- or 1-week-old spermatozoa stored at RT in Tesla-treated or Tesla-untreated ampoules. (g) Full-term development of embryos obtained from spermatozoa stored in Tesla-positive or Tesla-negative ampoules stored at RT for up to 3 months. Ampoule of FD spermatozoa with (h) silica gel or (i) deoxidizer. (j) The rate of blastocyst development from embryos derived from FD spermatozoa stored at RT for 3 weeks without any agent (W/O), with silica gel (Silica), deoxidizer (Deoxi), or both (Silica Deoxi). Different letters indicate statistically significant differences (P < 0.05). (k) Full-term development of embryos derived from FD spermatozoa stored at RT for up to 3 months with or without silica gel. Asterisk or different letters indicates statistically significant differences between samples (P < 0.05).